Promoting microalgal biofilm formation by crushed oyster shell-hydroxyapatite layer on micropatterned aluminum coating for heavy metal ions removal.

Microalgal biomass has shown inspiring potential for the heavy metal removal from wastewater, and forming microalgal biofilm is one of the sustainable methods for the microalgal biomass production. Here we report the formation of microalgal biofilm by accelerated colonization of typical algae Chlorella on thermal sprayed aluminum (Al) coatings with biologically modified surfaces. Micro-patterning surface treatment of the Al coatings promotes the attachment of Chlorella from 6.31 % to 17.51 %. Further enhanced algae attachment is achieved through liquid flame spraying a bioactive crushed oyster shell-hydroxyapatite (CaCO3-HA) composite top layer on the micropatterned coating, reaching 46.03-49.62 % of Chlorella attachment ratio after soaking in Chlorella suspension for 5 days. The rapidly formed microalgal biofilm shows an adsorption ratio of 95.43 % and 85.23 % for low concentration Zn2+ and Cu2+ in artificial seawater respectively within 3 days. Quick interaction has been realized between heavy metal ions and the negatively-charged extracellular polymeric substances (EPS) matrix existing in the biofilm. Fourier transform infrared spectroscopy (FTIR) results indicate that both carboxyl and phosphoryl groups of biofilms are crucial in the adsorption of Cu2+ and the adsorption of Zn2+ is due to the hydroxyl and phosphate groups. Meanwhile, the biofilm could act as a barrier to protect Chlorella against the attack of the heavy metal ions with relatively low concentrations in aqueous solution. The route of quick cultivating microalgal biofilm on marine structures through constructing biological layer on their surfaces would give insight into developing new techniques for removing low concentration heavy metal ions from water for environmental bioremediation.

Optimized production of full-length PCV2d virus-like particles in Escherichia coli: A cost-effective and high-yield approach for potential vaccine antigen development.

Porcine circovirus type 2 (PCV2) is a globally prevalent infectious pathogen affecting swine, with its capsid protein (Cap) being the sole structural protein critical for vaccine development. Prior research has demonstrated that PCV2 Cap proteins produced in Escherichia coli (E. coli) can form virus-like particles (VLPs) in vitro, and nuclear localization signal peptides (NLS) play a pivotal role in stabilizing PCV2 VLPs. Recently, PCV2d has emerged as an important strain within the PCV2 epidemic. In this study, we systematically optimized the PCV2d Cap protein and successfully produced intact PCV2d VLPs containing NLS using E. coli. The recombinant PCV2d Cap protein was purified through affinity chromatography, yielding 7.5 mg of recombinant protein per 100 ml of bacterial culture. We augmented the conventional buffer system with various substances such as arginine, ß-mercaptoethanol, glycerol, polyethylene glycol, and glutathione to promote VLP assembly. The recombinant PCV2d Cap self-assembled into VLPs approximately 20 nm in diameter, featuring uniform distribution and exceptional stability in the optimized buffer. We developed the vaccine and immunized pigs and mice, evaluating the immunogenicity of the PCV2d VLPs vaccine by measuring PCV2-IgG, IL-4, TNF-α, and IFN-γ levels, comparing them to commercial vaccines utilizing truncated PCV2 Cap antigens. The HE staining and immunohistochemical tests confirmed that the PCV2 VLPs vaccine offered robust protection. The results revealed that animals vaccinated with the PCV2d VLPs vaccine exhibited high levels of PCV2 antibodies, with TNF-α and IFN-γ levels rapidly increasing at 14 days post-immunization, which were higher than those observed in commercially available vaccines, particularly in the mouse trial. This could be due to the fact that full-length Cap proteins can assemble into more stable PCV2d VLPs in the assembling buffer. In conclusion, our produced PCV2d VLPs vaccine elicited stronger immune responses in pigs and mice compared to commercial vaccines. The PCV2d VLPs from this study serve as an excellent candidate vaccine antigen, providing insights for PCV2d vaccine research.

16S rRNA Gene Amplicon Sequencing of Gut Microbiota Affected by Four Probiotic Strains in Mice.

Probiotics, also referred to as "living microorganisms," are mostly present in the genitals and the guts of animals. They can increase an animal's immunity, aid in digestion and absorption, control gut microbiota, protect against sickness, and even fight cancer. However, the differences in the effects of different types of probiotics on host gut microbiota composition are still unclear. In this study, 21-day-old specific pathogen-free (SPF) mice were gavaged with Lactobacillus acidophilus (La), Lactiplantibacillus plantarum (Lp), Bacillus subtilis (Bs), Enterococcus faecalis (Ef), LB broth medium, and MRS broth medium. We sequenced 16S rRNA from fecal samples from each group 14 d after gavaging. According to the results, there were significant differences among the six groups of samples in Firmicutes, Bacteroidetes, Proteobacteria, Bacteroidetes, Actinobacteria, and Desferribacter (p < 0.01) at the phylum level. Lactobacillus, Erysipelaceae Clostridium, Bacteroides, Brautella, Trichospiraceae Clostridium, Verummicroaceae Ruminococcus, Ruminococcus, Prevotella, Shigella, and Clostridium Clostridium differed significantly at the genus level (p < 0.01). Four kinds of probiotic changes in the composition and structure of the gut microbiota in mice were observed, but they did not cause changes in the diversity of the gut microbiota. In conclusion, the use of different probiotics resulted in different changes in the gut microbiota of the mice, including genera that some probiotics decreased and genera that some pathogens increased. According to the results of this study, different probiotic strains have different effects on the gut microbiota of mice, which may provide new ideas for the mechanism of action and application of microecological agents.

Fabrication of Porous Aluminum Coating by Cored Wire Arc Spray for Anchoring Antifouling Hydrogel Layer.

Biofouling has been persisting as a worldwide problem due to the difficulties in finding efficient environment-friendly antifouling coatings for long-term applications. Developing novel coatings with desired antifouling properties has been one of the research goals for surface coating community. Recently hydrogel coating was proposed to serve as antifouling layer, for it offers the advantages of the ease of incorporating green biocides, and resisting attachment of microorganisms by its soft surface. Yet poor adhesion of the hydrogel on steel surfaces is a big concern. In this study, porous matrix aluminum coatings were fabricated by cored wire arc spray, and the sizes of the pores in the aluminum (Al) coatings were controlled by altering the size of the cored powder of sodium chloride. Silicone hydrogel was further deposited on the porous coating. The hydrogel penetrated into the open pores of the porous Al coatings, and the porous Al structure significantly enhanced the adhesion of the hydrogel. In addition, hydrogel coating exhibited very encouraging antifouling properties.

Signature Gene Identification of Cancer Occurrence and Pattern Recognition.

To identify signature genes for the pathogenesis of cancer, which provides a theoretical support for prevention and early diagnosis of cancer. The pattern recognition method was used to analyze the genome-wide gene expression data, which was collected from the The Cancer Genome Atlas (TCGA) database. For the transcription of invasive breast carcinoma, lung adenocarcinoma, lung squamous cell carcinoma, colon adenocarcinoma, renal clear-cell carcinoma, thyroid carcinoma, and hepatocellular carcinoma of the seven cancers, the signature genes were selected by means of a combination of statistical methods, such as correlation, t-test, confidence interval, etc. Modeling by artificial neural network model, the accuracy can be as high as 98% for the TCGA data and as high as 92% for the Gene Expression Omnibus (GEO) independent data, the recognition accuracy of stage I is more than 95%, which is higher compared with the previous study. The common genes emerging in five cancers were obtained from the signature genes of seven cancers, PID1, and SPTBN2. At the same time, we obtain three common pathways of cancer by using Kyoto Encyclopedia of Genes and Genomes' pathway analysis. A functional analysis of the pathways shows their close relationship at the level of gene regulation, which indicted that the identified signature genes play an important role in the pathogenesis of cancer and is very important for understanding the pathogenesis of cancer and the early diagnosis.

Long terminal repeat sequences from virulent and attenuated equine infectious anemia virus demonstrate distinct promoter activities.

In the early 1970s, the Chinese Equine Infectious Anemia Virus (EIAV) vaccine, EIAV(DLA), was developed through successive passages of a wild-type virulent virus (EIAV(L)) in donkeys in vivo and then in donkey macrophages in vitro. EIAV attenuation and cell tropism adaptation are associated with changes in both envelope and long terminal repeat (LTR). However, specific LTR changes during Chinese EIAV attenuation have not been demonstrated. In this study, we compared LTR sequences from both virulent and attenuated EIAV strains and documented the diversities of LTR sequence from in vivo and in vitro infections. We found that EIAV LTRs of virulent strains were homologous, while EIAV vaccine have variable LTRs. Interestingly, experimental inoculation of EIAV(DLA) into a horse resulted in a restriction of the LTR variation. Furthermore, LTRs from EIAV(DLA) showed higher Tat transactivated activity than LTRs from virulent strains. By using chimeric clones of wild-type LTR and vaccine LTR, the main difference of activity was mapped to the changes of R region, rather than U3 region.